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. 2023 Jan 6;24(2):1145. doi: 10.3390/ijms24021145

Table 1.

Selected methods for functional lncRNA analysis.

Method Reference
Determination of the intracellular lncRNA localization
Quantitative PCR (qPCR) [34]
RNA-fluorescent in situ hybridization (RNA-FISH) [35]
RNA-FISH combined with stochastic optical reconstruction microscopy (STORM) [36]
lncRNA labeling with aptamers linked to fluorescent tags [37,38]
lncRNA depletion or over-expression
Small interfering RNA (siRNA) silencing [78,79]
Short hairpin RNA (shRNA) silencing [80]
Antisense oligonucleotide (ASO) silencing [79]
CRISPR/Cas9 knock-out/knock-in [81]
The establishment of secondary and three-dimensional lncRNA structures
Shotgun secondary structure fragment analysis [40]
Solution-state nuclear magnetic resonance (NMR) spectroscopy [43,44]
Small-angle scattering (SAS) [45]
X-ray diffraction and cryo-electron microscopy [39]
Computational prediction [41,42,46]
The determination of lncRNA-protein interactions (LPIs)
Cross-linking immunoprecipitation (CLIP) [49,50]
Targets of RNA-binding proteins identified by editing (TRIBE) [51]
Digestion-optimized RNA immunoprecipitation cDNA library sequencing (DO-RIP-seq) [52]
RNA-affinity purification followed by mass spectrometry [53,54,55,56]
RNA-affinity purification followed by protein microarrays [57]
The isolation of target RNA molecules by biotinylated antisense probes [54,58]
The isolation of target RNA molecules by peptide nucleic acid oligomers [55]
HB-tag-based affinity RNA purification [56]
Chromatin isolation with RNA purification (ChIRP) [59,60]
RNA chromosome conformation capture (R3C) [61]
LPI computational prediction [72]
The biophysical characterization of quantitative and qualitative LPIs
Electrophoretic mobility shift assay (EMSA) [62]
Filter-binding assays [82]
Surface plasmon resonance [63,64]
The evaluation of the coding capacity of lncRNAs
Ribosome profiling [83]
Mass spectrometry [84]
Global translation initiation sequencing (GTI-seq) [73]